Mitigating Antibiotic Resistance
UFlorida iGEM 2021
TABLE OF
CONTENTS
01
Introduction
What is iGEM? Who are we?
02
Our Project Part 1
Our goals, background information and relevant
context
03
Our Project Part 2
The Science Stuff!
04
How you can get involved!
You can contribute to our project!
01 Introduction
What is iGem? Who are we?
iGEM stands for Internationally Genetically
Engineered Machine
What is iGEM?
PRIZES!
Best Foundational Advance Project
Best Environment Project
Best Energy Project
Best Information Processing Project
Best Hardware Project
Best Food & Nutrition Project
Best Diagnostics Project
Grand prizes!
Best Manufacturing Project
So… what is an iGEM team?
There are multiple components!
WET LAB
DRY LAB
HUMAN PRACTICES
WIKI
WET LAB
DRY LAB
HUMAN PRACTICES
WIKI
WHO ARE WE?
Scientists
Programmers
Engineers
College of Liberal Arts and Sciences
College of Engineering
Artists
02 Our Project Part 1
Background
Information
Our Goals
We wanted to tackle a problem...
So we just had several Zoom meetings where we brainstormed!
ANTIBIOTIC RESISTANCE!
So what is that exactly?
Antibiotic Resistance is...
Antibiotic Resistance threatens everyone...
Vocabulary alert!
Antibiotic Resistance
all
Antimicrobial Resistance
mean
Drug Resistance
the same thing!
WHERE DOES IT COME FROM?
Quick review on evolution...
1. Individuals in a population show variation
2. Variations can be inherited
3. Organisms have more offspring that can survive under normal circumstances
4. Variations that increase reproductive success will have a greater chance of being passed
on from generation to generation
5. A population will slowly change over time in response to the environment
So in terms of evolution...
Individuals in a population show variation not all bacteria have the same exact genes
Variations can be inherited bacteria with antibiotic resistance traits (at random!) can
reproduce
Organisms have more offspring that can survive under normal circumstances bacteria
exponentially grow, favorably genes will be duplicated rapidly
Variations that increase reproductive success will have a greater chance of being passed
on from generation to generation bacteria with antibiotic resistance genes are more
likely to survive and reproduce
A population will slowly change over time in response to the environment this is why
antibiotic resistance is a problem, there are resistant species emerging, not just
individual colonies
So, really, where is
this coming from?
ABR Sources
A brief history...
Select Germs Showing Resistance Over Time...
Antibiotic
Year
Released
Drug Target
Resistant Germ Identified
Year
Identified
Penicillin
1941
Bacterial cells with
peptidoglycan cell
walls
Penicillin
-resistant
Staphylococcus aureus
Penicillin
-resistant
Streptococcus pneumoniae
1942
1967
Methicillin
1960
Inhibiting bacterial
cell
-wall synthesis,
similar to Penicillin
Methicillin
-resistant
Staphylococcus aureus
1960
Fluconazole
1988
Inhibits fungal
enzyme required for
fungal cell wall
synthesis.
Fluconazole
-resistant Candida
1990
Vocabulary Alert!
Peptidoglycan
What is it???
How do successful antibiotics work and what is the basis of
resistance to them?
They are overusedPeople are misusing them
The Problem with Current Antibiotics….
Don’t stop when you “feel better.”
Finish the entire course.
Listen to your doctor!
Overprescribed, sometimes even
asked for when not needed.
They rely on similar mechanisms of action
Targeting the peptidoglycan wall
(for example)
Antibiotic Misuse results in….
Our Plan
Providing alternative selectable markers that do not rely on antibiotic resistance
genes
Alternative selectable markers are genes we can alter to produce obvious,
observable effects.
They allow us to determine which cells have been genetically altered and
which have not.
How to test?
Give bacteria a plasmid of interest with our DNA of interest (via conjugation)
Cells that have taken up DNA of interest (with an antibiotic resistance marker
on it) will be able to survive when plated on the respective antibiotic while
those who have not taken up the DNA of interest will die when plated on the
antibiotic
This is important in bacterial transformations because we want to see if a cell
took up a plasmid of interest
Our Plan continued....
We will be testing for carbon sources as positive selection markers
Sucrose, arabitol, and ribitol see if we can transform cells and give
them the ability to metabolize these sugars will be able to test for
transformants
03 Our Project Part - The Science Stuff!
Pseudomonas putida E. Coli
Bacteria we are using
Sugars we are working with...
Sucrose Arabitol Glucose Ribitol
Wet lab has
been busy...
31 2 3 4 5
7 8 9 10 11 12
14 15 16 17 18 19
21 22 23 24 25 26
28 29 30 31 1 2
130
6
13
20
27
Research
TestTestTest
Meeting
Lab Day
Lab Day Lab Day Lab Day
Lab Day Lab Day Lab Day
Lab Day
Lab Day
Lab Day
Our progress!
So far we have….
Made plasmids with each of the four sugar pathways
Made each plasmid with a backbone including antibiotic resistance
Prepared plates with our sugars and put our bacteria on their plates
with corresponding inducers
METHODOLOGY
and
EQUIPMENT
Venus has a beautiful
name and it’s hot
Jupiter is a bright object
in the night sky
Saturn is composed of
hydrogen and helium
Pipettes, Centrifuges, Autoclaves, Plates
PCR
Gel Electrophoresis
PCR
Polymerase Chain Reaction
So how does PCR
work?
GEL
ELECTROPHORESIS
Gel Electrophoresis
continued….
The end result!
EQUIPMENT
Just the basics….
Pipettes
Plates
Centrifuge
Autoclave
Just to wrap it all
together again
Why this is important….
HOW YOU CAN GET
INVOLVED!
We’re holding an infographic/meme competition!
Send us your best memes, artwork, or infographics related to iGEM,
synthetic biology, or antibiotic resistance, and we will credit you on our
iGEM wiki (if you want)!
If your work is selected for publishing, you can put on your resume that
you assisted with a University of Florida iGEM team project, and put our
names and wiki URL as a reference!
Top 3 winners get to contact us for advice related to UF, iGEM, science
research, and get a Zoom lab tour!
Rules: unlimited submissions, anything related to this presentation, and
make sure it is funny or artistic (whatever you are going for)
Submission Instructions
Email: ufigemteam@gmail.com
Subject line: RISE project
Send in highest resolution possible! We accept PNG, TIFF, JPG, PDF, etc.
Examples
CONCLUSIONS
Antibiotic Resistance is coming! Do your part to
complete your dose and listen to your doctor!!!
Science is fun! But complicated!
Keep iGEM and UF on your radar in the future!
Follow us on Instagram and don’t forget to send us
your submissions!
@uf.igem